Comparative Evaluation of GFP Expression in T98G (Human Brain Glioblastoma) cells after transfection by FuGENE®HD and Competitor Lipofectamine® 2000, 15,000 cells / well

 

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FuGENE®HD

Lipofectamine® 2000


Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5min
Complex formed in Sterile Deionized Water
Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20min

Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5min
Complex formed in OptiMEM®
Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20min

Reagent / DNA Ratio
0.4μl/0.1μg
8μl Complex per well
Complex Incubation 5min
Complex formed in Growth Media with 10% Fetal Bovine Serum
Reagent / DNA Ratio
0.5μl/0.2μg
50μl Complex per well
Complex Incubation 20min
Transfection with competitor reagent was performed according to manufacturer’s instructions

Protocol for T98G (Human Brain Glioblastoma) Cell Transfection by FuGENE HD with gWIZ™ GFP plasmid.

  1. Cell plating

    T98G cells were seeded from 90-100% confluent culture the day before transfection with the density 15,000 cells/well in 100 μl complete growth MEM medium supplemented with 1 mM Sodium Piruvate, 10 mM MEM non-essential Amino Acids Solution and 10% Fetal Bovine Serum.

  2. Complex preparation (per 20 wells)

    Tissue culture 96-round bottom well plates were used for complex preparation:

    1. Prepare 0.0125μg/μl gWIZ™ GFP DNA solution in OptiMEM®, sterile deionized water or growth media.
    2. Add 8 μl of reagent to 160 μl of DNA solution.
    3. Mix carefully by pipetting (15 times).
    4. Incubate 5 min at room temperature.
  3. Incubation

    Place the cells into CO2 incubator for 48 hours.

  4. Detection of GFP expression

    Monitor the transgene expression with fluorescent microscope or flowcytometer.